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Foot-and-mouth disease (FMD) is a highly contagious disease that infects cloven-hoofed animals, becoming a serious threat to livestock production and leading to significant economic losses. The re-occurring FMD outbreak in Indonesia was reported back in 2022, causing hundreds of cattle deaths. The immunogenic viral capsid VP1 protein has been extensively researched as a vaccine candidate despite the fact that the existing FMD vaccine uses an inactivated virus. The vp1 gene (648 bps) from FMD virus serotype O was integrated into pET-32b vector and transformed into Escherichia coli TOP10F’. The recombinant pET32b-VP1-1D plasmid was expressed in E. coli BL21(DE3), followed by N-terminal His tag purification. Protein profiles were determined with SDS-PAGE, showing the target protein at 33KDa. Five 6-week-old BALB/c mice were administered intraperitoneal injections of 50 μg and 100 μg protein, respectively, with two booster shots within two-week intervals. The immune response of polyclonal antibodies was tested using indirect ELISA, resulting in a high absorbance signal compared to non-immunized mice. Thus, the outcomes demonstrate that the VP1 recombinant protein from this study has potential as an immunogen in FMD vaccine development.